BIO Web of Conferences 9, 02034 (2017) DOI: 10.

1051/bioconf/20170902034
40th World Congress of Vine and Wine

Approaches to outline the aromatic profile of Kyoho wines from

South Korea
Enoch Yang, Foteini Drosou, Euthalia G. Dourtoglou, Archodoula Chatzilazarou, and Vassilis G. Dourtogloua

Department of Oenology and Beverage Technology, Faculty of Food Technology and Nutrition, Technological Educational Institute of
Athens, 12 Ag. Spyridonos St., Egaleo, 122 10 Athens, Greece

Abstract. Kyoho cultivar is a hybrid of V. vinifera and V. labrusca and is one of the major varieties cultivate
in South Korea. Kyoho grapes are characterized for their intense grape flavor and sour attributes. Korean
climatic conditions are different from worldwide wine producing countries. The summer season is hot and
dominated by heavy rains. Wines from Kyoho grapes are distinct for their intense grape flavor and the key
odors that characterize the wines are aromas of caramel and strawberry notes. The past few years efforts are
put to improve the quality and to advance the potential of the domestic wines. The objective of the present
study is to provide insight to the main aromatic compounds that form the aroma profile of Kyoho wines. For
this aim, Gas Chromatography was coupled with Mass Spectrometry for the separation and identification of
the compounds. Analysis allowed to identify the volatile compounds that form the aroma of the wine and to
detect the key odors of its characteristic aroma. (2,5-dimethyl-4-hydroxy-3(2H)-furanone) Furaneol
, which
is associated with caramel-like flavor, was one of the compounds that were detected along with ((3R)-hydroxy-
4,4-dimethyloxolan-2-one) pantolactone.

1. Introduction as well as their alcohol and flavor production [4].

The past years the grape industry in Korea has been
showing a positive trend in terms of increase area and
production volume. Korean grapes are mainly produced
for table use and only a small portion is used for
wine production. Owing to hot and rainy weathers in
the summer and cold conditions in winter, the climatic
conditions in Korea are unsuitable for the growth of
V.vinifera. [1] Thus, Korean wine makers try using
table grape from V. labrusca cultivars and their hybrids
to produce wines, as American grapes show stronger
adaptation to the adverse weather conditions [1, 2].
Main grapes growing in Korea are, ‘Campbell Early’
which occupies 74.3% of the total production area,
‘Kyoho’ which occupies 13.1% and ‘Muscat Bailey A’
which covers a small portion of 5.9% of the total area
cultivated for grape production [1, 3]. The majority of
Korean domestic wines are produced using the most
cultivated variety, that is Campbell Early. Though it has
the limitations of containing low levels of sugar, high malic
acid concentration and significant foxy aroma content that
affects negatively its preference [4]. Kyoho, the second
most cultivated grape, is a tetraploid hybrid of V. labrusca
and V. vinifera (‘Ishihara Wase’ × ‘Centennial’) [5–7] and
it is famous for its large-sized berry and for its intense
grape flavor [5]. Several studies have been conducted for
the improvement and optimization of the wines produced
from V. labrusca grapes. Hong & Park [4] analyzed the
diversity of indigenous yeasts isolated from Campbell
Early grapes. Furthermore, the identified yeast strains were
examined for their tolerance to potassium metabisulfite
a e-mail: vdourt@teiath.gr

c The Authors, published by EDP Sciences. This is an Open Access article distributed under the terms of the Creative Commons Attribution License 4.0
(http://creativecommons.org/licenses/by/4.0/).
Ahn et al. [8], examined the physiochemical differences
between V.vinifera and V. labrusca grapes by analyzing
their organic acid contents. Son et al. [9] used targeted
analysis of 1 H NMR to characterize and differentiate
between V. labrusca grapes and their wines. In their study,
Kyoho wines presented the highest levels of glycerol,
2,3-butanediol, succinate and alcohol compared to other
V. labrusca wines. Lee et al. [10] evaluated the sensory
attributes of a mixture of two different grape varieties,
Kyoho and Campbell Early and its chemical attributes
were determined and compared to a French Beaujolais
wine. Principal component analysis of the descriptive data,
showed high intensities of “ripe-fruit aroma”, “caramel
aroma” and “sour attributes for Kyoho wines.
In this study, the volatile compounds of Kyoho
wines were analyzed to determine which groups of
compounds consist the aroma profile of the wines and
which compounds contribute to the grape/caramel-like
odors of the wines.
2. Material and methods
2.1. Chemicals
Dichloromethane, chloroform, sodium chloride and anhy-
drous sodium sulphate were purchased from Chem Lab
(Zedelgem, Belgium). (2,5-dimethyl-4-hydroxy-3(2H)-
furanone) Furaneol
was purchased from Sigma-Aldrich
(Athens, Greece).
2.2. Samples
For the fulfillment of this study four monovarietal Kyoho
wines of two different produces were examined (ES and
BIO Web of Conferences 9, 02034 (2017)
40th World Congress of Vine and Wine
GS). From each producer two wine samples were provided
(ES1, ES2 and GS1, GS2, respectively). The grapes were
obtained and produced in Cheonan, Korea.
2.3. Volatile analysis
The extraction of volatile compounds was carried away by
Liquid-Liquid extraction (LLE) using dichloromethane as
elution sorbent. For the analysis of the volatile compounds
of the each examined wine, 50 mL of sample, 10 mL of
saturated NaCl solution and 50 mL of dichloromethane
were added in laboratory flask. The flask was stirred for
10 min at room temperature. Then, the sample was cen-
trifuged at 3500 rpm for 5 min for the separation of phases.
The supernatant was then extracted again using the same
volume of solvent for 10 min. The organic layers were then
washed with distilled water in a separation funnel. The
organic phase was dried over anhydrous sodium sulphate,
filtered and then condensed in a vacuum rotary evaporator.
Prior to instrumental analysis, the sample was recovered to
50 µL with Chloroform. For the injection 1 µL was used.
Each sample was analyzed in duplicate.
The identification of examined compounds was
made using their retention time and by compar-
ing their mass spectra with a spectral library of
known standard compounds. For the quantification of
(2,5-dimethyl-4-hydroxy-3(2H)-furanone) Furaneol
ex-
ternal calibration curves were prepared from standards. For
the quantification of ((3R)-hydroxy-4,4-dimethyloxolan-
2-one) Pantolactone and (2,5-dimethyl-4-methoxy-3(2H)-
furanone) Mesifuran, the area of the corresponding peaks
was normalized by that of the corresponding internal
standard. The quantification of the compounds was made
in Full Scan. The internal standard used for quantification
was 3-Octanol (3.0 mg/L). The GC peak area of
each compound was obtained from the ion extraction
chromatogram by selecting target ions for each one.
2.4. Gas Chromatography/ mass spectrometry
analysis
The apparatus used was an Agilent 6890 series GC System
(Agilent Technologies, USA), equipped with 5975C VL
MSD and a fused silica capillary column, 30m ×
0.32 mm i.d. × 0.25 µm film thickness (HP-5MS, Agilent
Technologies). 1.0 µL of sample was injected using a split
ratio of 100:1. The injector temperature was set at 250◦ C,
the carrier gas was helium at a flow rate of 1 mL/min,
and the oven temperature program was 50◦ C for 2.5 min,
increased to 180◦ C at 2.5◦ C /min, then to 230◦ C at 2◦ C
/min, then to 250◦ C at 6◦ C /min, then the temperature was
held at 250◦ C for 5 minutes, then increased to 270◦ C at
5◦ C/min and lastly it was held for 2 mins at 270◦ C. The
temperature of the transfer line was set at 280◦ C. The mass
spectrometer operated in the ionization mode (EI) at an
ionization voltage of 70 eV in the mass range 40–550 amu
and a manifold temperature of 270◦ C. All data was
recorded using the Turbomass 5.0 ChemStation software.
3. Results and discussion
3.1. Aroma volatiles composition by GC-MS
A total of 52 compounds were identified across four
Kyoho wine samples. The determined compounds are
2
DOI: 10.1051/bioconf/20170902034
summarized and classified by their functional groups in
Table 1. Among the determined compounds, the group
with the most compounds identified were higher alcohols
and esters. Altogether 16 alcohols, 15 esters, 7 fatty acids,
3 furans, 2 lactones and 3 miscellaneous compounds were
detected.
Higher alcohols, also known as fusel alcohols, are,
from a quantitative point of view, the most important
group of volatile compounds produced by yeast during
alcoholic fermentation. [11] Among the higher alcohols in
wines, compounds which exhibit the greatest abundance
are isoamyl alcohol 2-phenylethanols. 2-phenylethanol is
one of the most important phenol-derived higher alcohol
and it produces rose-like fragrances. [12–14]. Likewise,
Kyoho wines presented significant peaks for the above-
mentioned compounds.
Esters are mainly by-products of the alcoholic
fermentation and they are considered important to the
sensory properties of wines, as they contribute fruity
notes. Among the detected esters in the examined wines,
major compounds which are known to contribute to the
fruity character of a wine are isoamyl acetate, 2-phenyl
acetate, ethy hexanoate and ethyl decanoate. [13, 15] These
compounds were determined in all samples. Iso-butyl
acetate which has a strawberry-like odor was detected only
in one wine as shown in Table 1.
Fatty acids are mainly formed by yeasts during
alcoholic fermentation. Volatile fatty acids tend to have
a negative effect on wine aroma, contributing green
and cheese, fatty aromas [12, 13]. Some of the fatty
acids that were identified in the examined wines were
octanoic and decanoic acids. Free fatty acids occur
only as traces in grapes, but they are biosynthesized
during fermentation by yeasts and bacteria. Considering
the compounds’ unpleasant odors and similar ranges of
concentrations across wines, these acids are not considered
to be important contributors to the flavor of these wines
[10].
Among phenolics, Eugenol was detected which is
characterized as clove-like note. At usual concentrations,
eugenol adds a general spic note as well [16].
In the examined Kyoho wines two compounds
containing the 1-oxacycloalkan-2-one structure were
identified; γ -butyrolactone and pantolactone. Concentra-
tions of pantolactone were from 0.063 to 0.145 mg/L.
Pantolactones appear to be generated in grapes under
carbonic maceration, derived from amino or organic acids,
notably glutamic and succinic acids [14]. γ -Butyrolactone,
has been suggested that it contributes to the aroma
characteristics of Sherry wines, giving pleasant cotton,
peachy-like odors [11].
Contrast to major substances, minor classes can
contribute greatly to the flavor of the wine differentiating
each wine. Such classes are furans. Furans have very
sweet aromas and are normally classified in the same
group of aromas with give an odor of “burnt” sugar and
caramel. [17] Furaneol
, pantolactone and (2,5-dimethyl-
4-methoxy-3(2H)-furanone) mesifuran, compounds that
contribute to a sweet odor, were detected in all wines,
except for mesifuran which was not found in GS2. The
quantified concentrations of all three compounds are listed
in Table 2.
The quantities of mesifuran determined in the Kyoho
wines ranged between 0.046 to 0.066 mg/L. It is reported
BIO Web of Conferences 9, 02034 (2017) DOI: 10.1051/bioconf/20170902034
40th World Congress of Vine and Wine

Table 1. Volatile compounds detected in Kyoho wines.

Compound ES1 ES2 GS1 GS2 Odor description
Higher alcohols
1-Butanol – – – + Intoxicant alcoholb
Isobutanol – – – + Fusel, alcoholb
1-Pentanol – – + + Fusel alcohol, sweet balsamc
Iso-Amyl alcohol + + + + Harsh, fruity, bananaa, c
Prenol + + + + Fruity, green lavenderc
3-methyl-1-pentanol – + + + Alcohol, fusel, cognac, green fruityb, c
Isohexanol – + + + Nuttyc
1-Hexanol + + + + Green grassa
3-Hexen-1-ol + + + + Green leafyc
2-Ethyl-1-hexanol + + + + Mushroom, citrus floralb, c
1-Heptanol – – – + Fragrant plantb
Benzyl alcohol + + + + Floral, rosec
Phenylethyl alcohol + + + + Floral, rose, rose watera, c
Tyrosol + + + + Mild sweet, floral, fruityc
Homovanillyl alcohol + + + + N.f.
Tryptophol + + + + N.f.
Esters
Ethyl Lactate + + + + Solventa
Iso-Butyl acetate – – – + Strawberry, fruity, floweryb
Ethyl Butanoate + + + + Sour fruit, strawberryb
Isoamyl Acetate + + + + fresh banana
Isoamyl Lactate + + + + Sharp tart, buttery, butterscotchc
Ethyl Hexanoate + + + + Green apple, fruityb
Diethyl Succinate + + + + Light fruityb
Methyl vanillate + + + + Warm spicy, vanillac
Ethyl vanillate + + + + Phenolic, burnt, smokyc
Ethyl octanoate + + + + Melon, pineapplea, b
Phenethyl acetate + + + + Honey, pollen, rose b
Ethyl Decanoate + + + + Fruity, fatty, pleasantb
Succinic acid, ethyl 3-methylbutyl ester + + + + N.f.
Ethyl Dodecanoate + + – – Fruity, fatty, pleasantb
Ethyl palmiate + + + + Waxy, fruity, creamyc
Acids
Octanoic acid + + + + Rancid, harsh, cheesec
Decanoic acid + - + + Fatty, unpleasantc
Tetradecanoic acid + + + + N.f.
Hexadecanoic acid + + + + N.f.
Octadecanoic acid - + - - N.f.
Homovanillic acid + + + + N.f.
Homo-syringic acid + + + + N.f.
Phenols
Phenol, 2,5-di-tert-butyl- + + + + N.f.
Eugenol + + + + Clovec
Phenol, 2,6-dimethoxy-4-(2-propenyl) + + - - N.f.
4-((1E)-3-Hydroxy-1-propenyl)-2-methoxyphenol + + + + N.f.
Phenol, 2,4-bis(1,1-dimethylethyl)- + + + + N.f.
Furans
2-Furanmethanol - - + + Piquancy, bitterb
Furaneol
+ + + + Caramelb, c
Mesifuran + + + - Sweet, maple, fruityc
Lactones
Pantolactone + + + + Cotton candyc
γ −Butyrolactone + + + + Creamy, peachd
Miscellaneous
6R,9R-3-Oxo-α-ionol + + - + Spicyc
Limonene - - + + Lemon, orangea
3-Hydroxy-beta-damascone + + - + N.f.
N.f.: not found
a
Refers to Wang et al. (2016) [13]
b
Refers to Wang et al. (2017) [19]
c
Obtained from the Good Scents Company, http://www.thegoodscentscompany.com [20]
d
Refers to Pereira et al. (2014) [21].

3
BIO Web of Conferences 9, 02034 (2017)
40th World Congress of Vine and Wine
Table 2. Concentrations of quantified volatile compounds
(mg/L) a .
ES1 ES2 GS1 GS2
Furaneol
0.384 0.459 0.066 0.098
Mesifuran 0.046 0.063 0.066 N.d.
Pantolactone 0.098 0.094 0.063 0.145
N.d.: Not detected
a
Means are derived from duplicate determinations.
that mesifuran has been detected in substantial amounts in
V. labrusca hybrids often giving a caramel-like undertone
[18].
The determined concentration of Furaneol
ranged
between 0.066 to 0.459 mg/L. As stated by Rapp et al.
(1980) in [17], Furaneol
can be found in quantities
above 1mg/L in wines elaborated with hybrid grapes;
concentrations at which it produces a disagreeable
strawberry scent. In our case, although the examined
wines of our study did not present concentrations
above 1mg/L, the wines had a predominant odor of
strawberries and caramel-like notes, which can be
imparted to the presence of Fureaneol and Mesifuran.
Similar sensory attributes were described in the work of
Lee et al. [10]. A blend of Kyoho and Campbell Early
(V. labrusca varieties) wines were evaluated organolep-
tically; descriptive analysis showed high correlation for
grapiness, sweetness and sourness. Likewise, Jackson [16]
mentions that it is widely referred for V. labrusca grape
cultivars to possess intense strawberry odor, which is prob-
ably induced by furaneol
and tis methoxy derivatives.
Other wines in which Furaneol
imparts a significant
role are rose wines from Provence. Masson & Schneider,
[15] identified that the key compounds contributing to
the distinct aroma of Provence rose wines are derived
by the furanic compounds furaneol
and homofuraneol,
the C13 norisoprenoid (β-damascenone) and the two
volatile thiols 3-mercaptohexanol and 3-mercaptohexyl
acetate. In their study, the wines examined presented high
concentrations of furaneol
which indicate that furaneol

play a significant role in the aroma of rose wines. Similarly,
other authors in [13] affirm that the key compounds of
rose wines are ethyl esters, higher alcohol acetates (par-
ticularly isoamyl acetate and 2-phenyl acetate), furaneol

and the polyfunctional thiols 3-mercapto-1-hexanol and
3-mercaptohexyl acetate. In our study polyfunctional
thiols were not detected but it could be due to inadequate
sensitivity in the extraction procedure as the polyfunctional
thiols are present in extremely low concentrations [13].
Besides these compounds, β-damascenone is considered
to contribute significantly in the rose wine key odors
enhancing their fruity notes [15]. Taking into consideration
the fact that Kyoho grapes do not produce deep red wines,
but are weak in color as stated in [10], attempts to produce
rose wines could be encouraged, since they present similar
key odors to the Provence rose wines.
4. Conclusions
In summary, this study aimed to determine the volatile
compounds of Kyoho grapes. Specifically, this work
DOI: 10.1051/bioconf/20170902034
involved comprehensive evaluation of 4 Kyoho wines,
in which its sweet, strawberry, caramel like odor were
identified; these sweet aromas are characteristic of all
V. labrusca grapes even in their hybrid grapes showing
intense grape flavor, roses and strawberry like odors [14],
which are imparted by furans like furaneol
and its
methoxy derivatives.
This research has been co-funded by the European Union
(European Social Fund-ESF) and Greek national funds through
the Operational Program “Education and Lifelong Learning” of
the National Strategic Reference Framework (NSRF) – Research
Funding Program: ARCHIMEDES III. Investing in knowledge
society through the European Social Fund.
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